A novel approach is needed to overcome the deficiencies of protein production technology and ensure the produced enzymes fold into physiologically effective three-dimensional conformations. We discovered that incorporation of novel archaeal chaperones into a host-vector system and into in vitro enzymatic reactions helps both to recover proteins from inclusion bodies and to keep enzymes operationally active in vitro. This finding opens a way for novel protein folding technology both in vivo and in vitro. We propose to develop new host-vector systems, reagents and protocols based on archaeal chaperones for protein folding in vivo and in vitro. Archaeal chaperones exhibit folding activity for a broader range of substrates because archaeal species have evolved to occupy extreme ecological niches, which pose particular challenges to the problem of protein folding. In Phase 1, we will construct several host-vector systems to demonstrate the feasibility of our approach to produce aggregation-prone proteins in their native conformations with the help of archaeal chaperones. We will also develop simple assays to accelerate technology development, streamline problem management and enable quality control for new products and protocols. A robust protein production and folding technology plays a pivotal role in biotechnology innovation. It simplifies the multiple tasks of proteomics and encourages engineering new proteins for healthcare, environmental, and industrial purposes. PUBLIC HEALTH RELEVANCE: A robust and versatile protein folding technology will impact the healthcare related R&D programs at multiple levels. Most importantly, it will facilitate high-throughput production and purification of medically important proteins. It will also bring to a new level our ability to recover and analyze effective proteins in physiological three-dimensional conformations in vitro.